Synthetic biology approach for plant protection using dsRNA

Plant Biotechnology Journal
Annette NiehlManfred Heinlein

Abstract

Pathogens induce severe damages on cultivated plants and represent a serious threat to global food security. Emerging strategies for crop protection involve the external treatment of plants with double-stranded (ds)RNA to trigger RNA interference. However, applying this technology in greenhouses and fields depends on dsRNA quality, stability and efficient large-scale production. Using components of the bacteriophage phi6, we engineered a stable and accurate in vivo dsRNA production system in Pseudomonas syringae bacteria. Unlike other in vitro or in vivo dsRNA production systems that rely on DNA transcription and postsynthetic alignment of single-stranded RNA molecules, the phi6 system is based on the replication of dsRNA by an RNA-dependent RNA polymerase, thus allowing production of high-quality, long dsRNA molecules. The phi6 replication complex was reprogrammed to multiply dsRNA sequences homologous to tobacco mosaic virus (TMV) by replacing the coding regions within two of the three phi6 genome segments with TMV sequences and introduction of these constructs into P. syringae together with the third phi6 segment, which encodes the components of the phi6 replication complex. The stable production of TMV dsRNA was achieved by...Continue Reading

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Citations

May 11, 2019·International Journal of Molecular Sciences·Alexandra S Dubrovina, Konstantin V Kiselev
Aug 23, 2019·Nucleic Acids Research·Selma Gago-ZachertSven-Erik Behrens
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Sep 28, 2021·Plant Biology·V OberemokR Kamenetsky-Goldstein

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Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

ImageJ

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